Haemophilus outer membrane protein

Abstract

Purified and isolated nucleic acid from specific strains of Haemophilus influenzae is provided which encodes at least a portion of the D15 outer membrane protein of Haemophilus. The nucleic acid is used to produce peptides, polypeptides and proteins free of contaminant associated with Haemophilus for purposes of diagnosis and medical treatment. Furthermore, the nucleic acid may be used in the diagnosis of Haemophilus infection. Antisera obtained following immunization with the nucleic acid D15 outer membrane protein or peptides also may be used for the purpose of diagnosis and medical treatment.

What we claim is:  
     
       1. An immunogenic peptide consisting of an amnio acid sequence which is that of a portion only of the amino acid sequence of a purified and isolated D15 outer membrane protein having a molecular weight of about 80 kDa, said peptide being selected from the group consisting of peptides consisting of SEQ ID NOS: 14-38, 40, 41 and 45-49. 
     
     
       2. An immunogenic composition, comprising at least one immunologically-active component which is selected from the group consisting of: 
       (a) a purified and isolated  Haemophilus influenzae  D15 outer membrane protein having a molecular weight as determined by SDS-PAGE of about 80 kDA and selected from the group consisting of:  
       (i) an amino acid sequence consisting of SEQ ID NOS: 2, 4, 6, 8 or 10;  
       (ii) an amino acid sequence encoded by DNA sequence consisting of SEQ ID NOS: 1, 3, 5, 7 or 9; and  
       (iii) an amino acid sequence encoded by a DNA sequence consisting of a consensus DNA sequence consisting of SEQ ID NO: 55;  
       (b) an immunogenic synthetic peptide consisting of an amino acid sequence which is that of a portion only of the amino acid sequence of a purified and isolated D15 outer membrane protein, said peptide being selected from the group consisting of peptides consisting of SEQ ID Nos. 14-36, 40, 41 and 45-49; and  
       (c) a chimeric molecule comprising the protein in (a) or the peptide in  
       (b) linked to a foreign polypeptide or protein or to a polysaccharide; and a physiologically-acceptable carrier therefor.  
     
     
       3. The immunogenic composition of claim  2  formulated as a vaccine for in vivo administration to protect against diseases caused by Haemophilus. 
     
     
       4. The immunogenic composition of claim  3  formulated as a microparticle preparation, capsule preparation or liposome preparation. 
     
     
       5. The immunogenic composition of claim  3  in combination with a targeting molecule for delivery to specific cells of the immune system or to mucosal surfaces. 
     
     
       6. A method of inducing protection against disease caused by Haemophilus, comprising administering to a subject an effective amount of the immunogenic composition of claim  2  to provide protection against Haemophilus disease. 
     
     
       7. A chimeric molecule, comprising: 
       (a) a purified and isolated D15 outer membrane protein having a molecular weight of about 80 kDa, wherein said outer membrane protein is selected from the group consisting of:  
       (i) an amino acid sequence consisting of SEQ ID NOS: 2, 4, 6, 8 or 10;  
       (ii) an amino acid sequence encoded by a DNA sequence consisting of SEQ ID NOS: 1, 3, 5, 7 or 9; and  
       (iii) an amino acid sequence encoded by a DNA sequence consisting of a consensus DNA sequence consisting of SEQ ID NO: 55 and  
       (b) an immunogenic synthetic peptide consisting of an amino acid sequence which is that of a portion only of the amino acid sequence of a purified and isolated D15 outer membrane protein having a molecular weight of about 80 kDa, said peptide being selected from the group consisting of peptides consisting of SEQ ID NOS: 14-38, 40, 41 and 45-49 linked to a foreign polypeptide or protein or to a polysaccharide.  
     
     
       8. The chimeric molecule of claim  7  wherein said D15 outer membrane protein is linked to a foreign polypeptide or protein which comprises a surface protein or peptide from a pathogenic bacteria. 
     
     
       9. The chimeric molecule of claim  8  wherein said foreign polypeptide or protein comprises a P1, P2, or P6 outer membrane of  H. influenzae.    
     
     
       10. The chimeric molecule of claim  7  wherein said D15 outer membrane protein is linked to a polysaccharide which comprises a PRP molecule from  H. influenzae.    
     
     
       11. A purified and isolated  Haemophilus influenzae  D15 outer membrane protein having a molecular weight as determined by SDS-PAGE of about 80 kDa and consisting of an amino acid sequence which is selected from the group consisting of: 
       (a) an amino acid sequence consisting of SEQ ID NOS: 2, 4, 6, 8 or 10;  
       (b) an amino acid sequence encoded by a DNA sequence consisting of SEQ ID NOS: 1, 3, 5, 7 or 9; and  
       (c) an amino acid sequence encoded by a DNA sequence consisting of a consensus DNA sequence consisting of SEQ ID NO: 55.  
     
     
       12. The protein of claim  11  wherein the  Haemophilus influenzae  is a  Haemophilus influenzae  type b strain selected from the group consisting of Ca, MinnA and Eagan strains. 
     
     
       13. The protein of claim  11  wherein the  Haemophilus influenzae  is a non-typeable  Haemophilus influenzae  strain selected from the group consisting of PAK12085 and SB33 strains. 
     
     
       14. A purified and isolated  Haemophilus influenzae  D15 outer membrane protein consisting of an amino acid sequence which has SEQ ID NO: 8 or consisting of an amino acid sequence which is encoded by a nucleic acid molecule which has SEQ ID NO:7.